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Effect of HIF-1α and its mechanism on invasion and metastasis of esophageal carcinoma Eca109 cells in vitro and in vivo

JING Shao-wu1,WANG Ya-di2,ZHENG Ming-min2,SUN Guo-gui1,LIU Qing1,CHENG Yun-jie1,YANG Cong-rong1 1Department of Radiotherapy,Fourth Hospital of Hebei Medical University,Shijiazhuang 050011;2Department of Radiotherapy,The Military General Hospital of Beijing PLA,Beijing 100700,China  
AIM:To investigate the effects of HIF-1α by RNAi on invasion and metastasis of esophageal carcinoma Eca109 cells in vitro and in vivo,in order to explore its probable mechanism.METHODS:CoCl2 was used to mimic tumor hypoxic microenvironment.mRNA and protein levels of HIF-1α,E-cadherin and MMP-2 under hypoxia were detected by RT-PCR and immunohistochemistry.The effects of silencing HIF-1α by RNAi on HIF-1α,E-cadherin and MMP-2 mRNA were detected by RT-PCR.The effect of RNAi on invasion and metastasis were tested by cell scratch assay and transwell chambers.The Eca109-implanted nude mouse model was established,and the effects of HIF-1αon tumor growth and lymphoid node metastasis were observed.The expressions of HIF-1α,E-cadherin and MMP-2 in transplanted tumors were detected by Western blot,and the effects of HIF-1α on tumor growth,invasion and metastasis in vitro and in vivo were analyzed.RESULTS:Hypoxia up-regulated HIF-1α protein,mRNA and protein levels of E-cadherin down-regulated,and MMP-2 up-regulated,while had no effect on HIF-1α mRNA.RNAi could silencing HIF-1α effectively,and inhibited E-cadherin or MMP-2 decreased or increased,respectively.The migration and the number of invading cells decreased(P0.05) after silencing HIF-1α by RNAi.The tumor volume was much smaller,lymph node metastasis rate lower as well in vivo(P0.05).CONCLUSION:Via its effects on E-cadherin and MMP-2,HIF-1α regulate the growth,invasion and metastasis of Eca109 cells in vitro and in vivo.
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